KR101672804B1 - Tube for fully automatic centrifuge - Google Patents

Abstract

The present invention relates to a specific tube for a fully automatic centrifuge that can be applied to separate all biosamples. More particularly, the present invention relates to a method for the treatment and / or prophylaxis of a disease, such as a buffy coat layer comprising mesenchymal and hematopoietic stem cells, blood red blood cells, platelet rich plasma (PRP) Which can be quickly and easily separated, and which can be applied to portable, fully automatic centrifuges. It is also about a particular tube that can be used in a typical centrifuge.

 Stem cells, buffy coat, whole automatic centrifugation, pre-automatic sampling, tubes,

Description

A tube for a fully automatic centrifuge that can be applied to separate all bio samples.

The present invention relates to a specific tube for a fully automatic centrifuge that can be applied to separate all biosamples. More particularly, the present invention relates to a method for the treatment and / or prophylaxis of a disease, such as a buffy coat layer comprising mesenchymal and hematopoietic stem cells, blood red blood cells, platelet rich plasma (PRP) To a centrifugal separator which can be quickly and easily separated and which is also portable. It is also about a particular tube that can be used in a typical centrifuge.

Stem cells are 'undifferentiated' cells that have the ability to differentiate into various types of body tissues. They are collectively referred to as cells that have the potential to differentiate into various tissue cells when appropriate conditions are met. Cells and adult stem cells.

Embryonic Stem Cell is an embryonic stem cell of less than 14 days after fertilization. It refers to a cell that has the potential to differentiate into all the cells and tissues that constitute the human body in the future, whereas Adult Stem cell ) Refers to primordial cells that have been extracted from cord blood and umbilical cord blood (adult blood) and blood, and before they are differentiated into cells of specific organs such as bone, liver, and blood.

Adult stem cells are capable of differentiating into hematopoietic stem cells capable of differentiating into blood cells, tissue cells such as bones, cartilage, muscles, and nerves, and thus mesenchymal stem cells (MSCs) ) And neural stem cells capable of differentiating into neurons. These stem cells are not only the liver, epidermis, pancreas, bone marrow, but also dental pulp, peridontal ligament, Can be widely obtained at each of the regions. In particular, umbilical cord blood (umbilical cord blood) contains a large amount of hematopoietic stem cells, bone marrow cells found in the bone marrow cells, hematopoietic stem cells and hematopoietic stem cells can produce a variety of stem cells.

Although these adult stem cells are difficult to separate and proliferate, they are highly likely to be used in the field of regenerative medicine, such as regenerating damaged tissues, because they are highly likely to differentiate into specific organs. For example, skin lesions caused by diseases such as wounds or ulcers are likely to leave scars due to imperfect regeneration of early skin tissues or abnormal arrangement of collagen fibers after treatment. However, when stem cells are applied to such treatment, The occurrence of scars can be minimized. In fact, when stem cells are injected into damaged skin tissue, it has been studied that these stem cells contribute to the regenerated skin cell populations (Fathke et al., Stem Cells 2004; 22: 812-822).

In addition, in the case of implant treatment, stem cells can be used for regeneration steps such as root and associated periodontal tissues, thereby improving the surgical effect and shortening the procedure period. In fact, it has been studied that stem cells from dental and periodontal ligaments contribute to the creation of root / peridontal tissues complexes that can support crowns (Sonoyama W. et al, PLOS ONE, Issue 1, December 2006, e79). In addition, stem cell-based methods can be widely used in the recovery and regeneration of various treatments and procedures.

Meanwhile, in using stem cells for the above-mentioned treatment, if stem cells can be directly extracted from their own body and used directly for treatment, it will be the simplest and easiest way to utilize stem cells without side effects. Therefore, studies are being conducted to isolate stem cells for practical treatment and various analyzes.

For example, U. S. Patent Application Publication No. US2007 / 0265558 (published on December 15, 2007) discloses a series of methods for separating a buffy coat layer containing stem cells after blood is centrifuged . However, since the buffy coat layer accounts for only a fraction of 1% of the whole blood, a large amount of blood is required to obtain as many stem cells as necessary. Therefore, it is difficult to apply the above-mentioned smoke layer in a routine treatment process, and the separation process is also complicated, so that there is a problem in applying it to a treatment easily and quickly. In addition, it can be seen that it is difficult to use the above method to make a device for easily separating stem cells due to the amount of a large number of samples and the separation method comprising several steps.

In addition, US Patent Publication No. US2007 / 0092971 (published on March 26, 2007) discloses a separation tube capable of separating a buffy coat layer from blood, it has a complicated structure for separating the buffy coat layer. Therefore, it is difficult to use it easily and conveniently in daily treatment and operation.

Thus, a very small amount of specific layer such as a buffy coat layer containing stem cells can be selectively and easily obtained from a very small amount of blood or other samples of several to several tens of milliliters, It has been urgently required to study a separation tube which can be easily carried by a device and can also be used for military purposes (which can also be used outdoors).

It is an object of the present invention to apply to the separation of all biosamples, particularly buffy coat layers including mesenchymal and hematopoietic stem cells from blood, fat and bone marrow, red blood cells, And a platelet rich plasma (PRP) can be rapidly and easily separated.

It is another object of the present invention to provide a centrifuge tube that can be applied to a portable centrifuge.

It is another object of the present invention to provide a centrifuge tube that can be used in a general centrifuge.

The above object of the present invention is achieved by a centrifuge tube for a centrifuge comprising a first constituent part, a second constituent part and a third constituent part, wherein the diameter of the first constituent part and the third constituent part is larger than the diameter of the second constituent part . ≪ / RTI >

It is still another object of the present invention to provide a tube for a centrifuge, wherein the diameter of the second component is 1 mm or more and 50 mm or less.

It is still another object of the present invention to provide a tube for a centrifuge, wherein the length of the second component is 10 mm or more and 50 mm or less.

By using the tube for a centrifuge of the present invention, an extremely small amount of the sample can be selectively and quickly separated from a small amount of sample. In particular, since a buffy coat containing stem cells can be easily separated from a small amount of blood, fat, and bone marrow, such as several to several tens of milliliters, It is possible to shorten the period of regeneration and recovery and increase the efficiency thereof.

For example, in the case of implant surgery, dental pulp stem cells and peridontal ligament stem cells can be separated from the blood, fat and bone marrow from the procedure to shorten the procedure time In addition, the side effects can be minimized, and patients with poor alveolar bone that can not be implanted can help supplement alveolar bone.

When the biosample is separated using the tube, high reproducibility and pureness of a specific component can be obtained by using a sample of the same volume and the same volume at all times.

In addition, it is possible to separate and utilize a membrane layer containing stem cells from blood, fat and bone marrow in an urgent situation such as a military operation, and it can be applied to a centrifuge device which can be carried by a small volume Do. In addition, it can be used in practically all therapeutic processes such as hair treatment and skin treatment, analysis and research.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

1, the centrifugal separator tube of the present invention comprises a first component 11, a second component 12 and a third component 13 as shown in FIG. 1, and the first component 11 and the third component And the diameter of the part (13) is larger than the diameter of the second component (12). By making the diameter of the second component thinner than that of the first component and the third component, it was difficult to separate the tube because it was too thin due to the small amount when the tube had only one diameter. It can be extended easily to separate.

If the sample is blood, the blood is injected into a tube containing ficoll and centrifuged. After centrifugation, the blood is largely divided into a plasma layer, a buffy coat layer, a red blood cell (RBC) ) Layers. The plasma layer is formed in the first constituent part 11, the soft film layer is formed in the second constituent part 12, and the RBC layer is formed in the third constituent part 13 .

The plasma layer is a liquid component of the blood, accounting for about 55% of the whole blood sample. The buffy coat layer includes mononuclear cells including leukocytes, platelets, stem cells, etc. Lt; RTI ID = 0.0 > 1% < / RTI > of the whole blood sample. Smoke layers are usually used to extract DNA in the blood of mammals, usually with a slightly white color, but occasionally green. The RBC layer accounts for approximately 45% of the whole blood sample and contains most of the red blood cells along with the ficoll component.

In this case, when the general tube is used, it is possible to easily separate the layer, which is too thin and impossible to separate. Therefore, by using the tube of the above structure, the smoke layer can be effectively separated from the sample having a volume of 10 ml or less. At this time, one or more septums 14 may be formed on the wall surface of the second constituent part in order to directly separate the smoke layer located in the second constituent part, and a syringe or the like may be formed through the septum As shown in FIG. In this case, it is possible to use the separated polishing layer as it is in the human body by using a syringe containing a buffer containing an antibiotic in the separation step, and it is possible to use stem cells used as an ampoule from the syringe used in the separation process It is possible to store the formed film layer. On the other hand, a fixing device may be formed to fix the tube on one surface of the tube.

In a preferred embodiment of the present invention, the diameter of the second constituent part of the tube is preferably 1 mm or more and 50 mm or less, and a septum is formed on the second constituent part, and a syringe, The smoke layer can be separated immediately.

Also, in a preferred embodiment of the present invention, the length of the second component of the tube is preferably 10 mm or more and 50 mm or less, and if it is smaller, it may be difficult to separate all the smoke layer into the second component, If the length is too long, the overall length may be too long and inefficient. The length of the entire tube is preferably 100 to 200 mm.

On the other hand, the inner wall surface of the tube may be a plastic tube or a glass tube whose inner wall surface is coated with a glass material to prevent the sample from sticking to the wall surface when hydrophobic constituent materials are contained, such as blood. Further, it is preferable that the tube is made of a transparent material for spectroscopic classification of each centrifuged component.

Further, in applying the centrifuge tube to a centrifugal separator, a part to be separated can be automatically recognized and separated by using a spectroscopic device for recognizing and detecting centrifugal components. In addition, the centrifuge tube may be used together with a device such as a CCD camera (Charge Couple Device) and an image analyzer in order to recognize and detect the centrifuged components.

On the other hand, a fixing device 15 such as concavo-convex or magnetic material may be formed on the centrifuge tube so that the surface contacting the centrifuge tube with the sample extractor or the sample recognition device is constant.

It will be understood by those skilled in the art that the present invention is not limited to the specific embodiments and descriptions described above and that those skilled in the art, It will be understood that various modifications may be made without departing from the scope of the present invention.

1 is a cross-sectional view of a centrifuge tube of the present invention.

FIG. 2 is a conceptual diagram showing a sample taken from a diaphragm of a tube for a centrifuge of the present invention. FIG.

DESCRIPTION OF THE REFERENCE SYMBOLS USED IN THE DRAWINGS

11: tube first component 12: tube second component

13: tube third component 14: diaphragm

15: Fixing device

Claims (13)

A first component (11); A third component 13; And A second component (12) connecting the first component (11) and the third component (11) (13); Lt; / RTI > The diameter of the second constituent part 12 is 1 mm or more and 50 mm or less, The diameter of the first component 11 and the third component 13 is larger than the diameter of the second component 12, The length of the second constituent part 12 is 10 mm or more and 50 mm or less in the entire length of 100 to 200 mm, A septum 14 is formed on the side surface of the second component portion 12 to separate the smoke film layer, A plasma layer is formed on the first constituent part 11 and a second constituent part 12 is formed on the third constituent part 13 during centrifugation of blood, fat and bone marrow, Lt; RTI ID = 0.0 > RBC < / RTI > delete delete delete delete The centrifuge tube according to claim 1, wherein ficoll is used as a medium for centrifugation. The centrifuge tube according to claim 1, wherein the inner wall surface is made of plastic or a glass material coated with a glass material. delete The centrifuge tube according to claim 1, wherein the septum (14) is at least one of the septum (14). The centrifuge tube according to claim 1, wherein the centrifuge tube is made of a transparent material for spectroscopic classification of each centrifuged component. The centrifuge tube according to claim 1, characterized in that it is used together with a spectroscopic device for recognizing and detecting each centrifuged component. The centrifuge tube according to claim 1, characterized in that it is used together with a CCD camera (Charge Couple Device) and an image analyzer to recognize and detect each component centrifuged. The centrifuge tube according to claim 1, wherein a fixing device (15) is formed to make the surface of the sample extractor or the sample recognition detector constant.

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